Shi Hua, Fan Xiaochun, Ni Zhuoyu, Lis John T
RNA. 2002 Nov;8(11):1461-70. doi: 10.1017/s1355838202029941.
Iterative cycles of in vitro selection and amplification allow rare functional nucleic acid molecules, aptamers, to be isolated from large sequence pools. Here we present an analysis of the progression of a selection experiment that simultaneously yielded two families of RNA aptamers against two disparate targets: the intended target protein (B52/SRp55) and the partitioning matrix. We tracked the sequence abundance and binding activity to reveal the enrichment of the aptamers through successive generations of selected pools. The two aptamer families showed distinct trajectories of evolution, as did members within a single family. We also developed a method to control the relative abundance of an aptamer family in selected pools. This method, involving specific ribonuclease digestion, can be used to reduce the background selection for aptamers that bind the matrix. Additionally, it can be used to isolate a full spectrum of aptamers in a sequential and exhaustive manner for all the different targets in a mixture.
体外选择和扩增的迭代循环使得能够从大的序列库中分离出罕见的功能性核酸分子——适配体。在此，我们对一个选择实验的进程进行了分析，该实验同时产生了针对两个不同靶标的两个RNA适配体家族：预期的靶蛋白（B52/SRp55）和分配基质。我们追踪了序列丰度和结合活性，以揭示经过连续几代选择库后适配体的富集情况。这两个适配体家族显示出不同的进化轨迹，单个家族内的成员也是如此。我们还开发了一种方法来控制选择库中适配体家族的相对丰度。这种方法涉及特异性核糖核酸酶消化，可用于减少与基质结合的适配体的背景选择。此外，它可用于以连续且详尽的方式分离混合物中所有不同靶标的全谱适配体。
